Methods for improved poxvirus yields
AssigneeArizona Board of Regents on behalf of Arizona State University
InventorsDouglas Grant McFadden, Mohammed Masmudur Rahman
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Abstract
A method of producing poxviruses at an increased growth rate and/or progeny virus titer in cells, the method including: contacting a host cell with an effective amount of nucleocytoplasmic transport inhibitor; contacting the host cell with a poxvirus of interest under conditions that permit the poxvirus of interest to adsorb to the surface of the host cell; and culturing the host cell to produce progeny poxvirus of interest.
Claims12
A method of producing a poxvirus at an increased growth rate or titer, the method comprising:
- contacting a host cell with a nucleocytoplasmic transport inhibitor at a concentration of at least 0.01 μM;
- wherein the host cell is a transformed cell;
- contacting the host cell with a poxvirus, wherein the host cell is contacted with the poxvirus at a multiplicity of infection (MOI) less than one;
- and
- culturing the host cell to produce progeny of the poxvirus at an increased growth rate compared to the growth rate in identical conditions lacking the nucleocytoplasmic transport inhibitor or an at least 3-fold increase in titer compared to that produced in identical conditions lacking the nucleocytoplasmic transport inhibitor,
- wherein the nucleocytoplasmic transport inhibitor is one or more selected from the group consisting of:
- Leptomycin A, Leptomycin B, Ratjadone A, Ratjadone B, Ratjadone C, Ratjadone D, and Anguinomycin A;
- and
- wherein the poxvirus is myxoma virus.
A method for replicating a leporipoxvirus, the method comprising:
- exposing a host cell to a nucleocytoplasmic transport inhibitor at a concentration of at least 0.01 μM;
- wherein the host cell is a transformed cell;
- infecting the host cell with a leporipoxvirus at a multiplicity of infection (MOI) less than 1, thereby generating an infected host cell;
- and
- incubating the infected host cell in a culture medium under conditions that allow for replication of the leporipoxvirus,
- wherein the rate of replication is greater than the rate of replication in identical conditions lacking the nucleocytoplasmic transport inhibitor or an at least 3-fold increase in titer compared to that produced in identical conditions lacking the nucleocytoplasmic transport inhibitor
- wherein the nucleocytoplasmic transport inhibitor is one or more selected from the group consisting of:
- Leptomycin A, Leptomycin B, Ratjadone A, Ratjadone B, Ratjadone C, Ratjadone D, and Anguinomycin A;
- and
- wherein the leporipoxvirus is a myxoma virus.
Dependencies are inferred from the claim text.